Proton-translocating nicotinamide nucleotide transhydrogenase is located in the mitochondrial inner membrane and catalyzes the reduction of NADP(+) by NADH to NADPH and NAD(+). The present investigation describes the expression of the transhydrogenase gene in various mouse organs, subsections of the human brain and Caenorhabditis elegans. In the mouse, the expression was highest in heart tissue (100%) followed by kidney (64%), testis (52%), adrenal gland (41%), liver (35%), pancreas (34%), bladder (26%), lung (25%), ovary (21%) and brain (14%). The expression in brain tissue was further investigated in the human brain which showed a distribution that apparently varied as a function of neuronal density, a result that was supported by estimations of expression in C. elegans using Green Fluorescent Protein (GFP) controlled by the transhydrogenase promoter. GFP-expressing C. elegans lines showed a clear concentration of fluorescence to the gut, the pharyngeal-intestinal valve and certain neurons. It is concluded that the transhydrogenase gene is expressed to various extents in all cell types in mouse, human and C. elegans.